George C Tsokos
Beth Israel Deaconess Medical Center
Project start date: 2001-04-01
Project end date: 2012-12-31
Sponsored Links Excellgen http://Excellgen.com
George C Tsokos, Professor Of Medicine
Henry M. Jackson Fdn For The Adv Mil/med Advancement Of Military Medicine, Inc. Rockville, Md 20852
Grant 5R01AI049954-05 from National Institute Of Allergy And Infectious Diseases IRG: GMA
Abstract: Systemic lupus erythematosus (SLE) is an autoimmune disorder of indeterminate etiology characterized by profound T cell effector dysfunctions including defective interleukin-2 (IL-2) production. The fundamental underlying molecular mechanisms remain largely unknown. This laboratory has considered that deficient production of IL-2 represents decreased transcription of the IL-2 gene and has undertaken a systematic analysis to investigate aberrations of transcription factors that bind to the IL-2 promoter and cause defective IL-2 gene transcription. The presented preliminary studies show that IL-2 production by SLE T cells is decreased and the activity of two activators, nuclear factor (NF)-kappaB and activating protein (AP)-1, is decreased, whereas the activity of the repressor cAMP responsive element modulator (CREM) is increased. The latter binds to the -180 (-164/-189) site of the IL-2 promoter and suppresses gene transcription. This information suggests that the decreased IL-2 production in human SLE is the result of transcriptional repression mediated by the increased expression of CREM and/or the decreased activity of activators. This hypothesis will be addressed logically in experiments planned in 3 specific aims, which will 1) Establish in cross-sectional and longitudinal studies that the increased expression of CREM in human SLE subjects is disease-specific and disease activity-independent and that it relates directly with the decreased production of IL-2. 2) Determine how CREM suppresses the production of IL-2 in human SLE T cells by studying its ability to interact with other transcription factors on the IL-2 promoter. 3) Investigate the mechanisms that are involved in the increased expression of CREM in SLE T cells. Defective IL-2 production contributes to increased infection-related morbidity and mortality rates in patients with SLE. The proposed studies will contribute to our understanding of the origin of a major clinical problem.
Keywords: T lymphocyte, genetic transcription, immunogenetics, interleukin 2, molecular pathology, systemic lupus erythematosus, anergy, messenger RNA, transcription factor, human subject, patient oriented research, tissue /cell culture, transfection
Project start date: 2002-04-15
Project end date: 2006-08-31
5R01AI049954-05 (2006): $253256
5R01AI049954-04 (2005): $259350
5R01AI049954-03 (2004): $259350
5R01AI049954-02 (2003): $259350
5R01AI049954-09 (2010): $382500
Grants awarded to George C Tsokos
TRAINING PROGRAM IN SYSTEMIC AUTOIMMUNITY
George C Tsokos, Professor Of Medicine
Beth Israel Deaconess Medical Center, Boston, Ma 02215
Grant 5T32AI074549-03 from National Institute Of Allergy And Infectious Diseases
Abstract: This is a new proposal of a training program in systemic autoimmunity at the Beth Israel Medical Deaconess Medical Center (BIDMC), Harvard Medical School (HMS). It requests support for two trainees each year who will spend at least two years in the program. Trainees will enter the program through one of the following tracks Physicians who seek training in rheumatology or nephrology, physicians who enter other clinical fellowship programs including neurology and dermatology, physicians who have completed clinical training in a field relevant to systemic autoimmune diseases and PhDs with background in immunology, biochemistry and cell/molecular biology. The major criterion for selection will be evidence of commitment to the study of systemic autoimmune diseases. Through this training physicians will develop skills in immunology and biology/biochemistry, whereas PhDs will become familiar with clinical issues pertinent to the study of systemic autoimmune diseases. Research activities will span studies in human and animal T cells, B cells, NKT cells, macrophages, mast cells, costimulation, immune cell signaling, immune tolerance, control of cytokine expression and function, pertinent aspects of regional immunology and biology (kidney, skin and nervous system), human genetics, immunodeficiency, complement and immunomodulation. The chief method of instruction will involve personal involvement in a program of basic research under the close supervision of a senior scientist. Moreover, a structured program of didactic courses in research seminars, journal clubs, research presentations in national and international meetings and grant writing is in place. The primary facilities for training are at BIDMC and HMS-affiliated institutions. Graduating trainees will be able to plan, seek funding, and execute cutting edge research in systemic autoimmune diseases and will be properly tooled to develop new diagnostics and therapeutics. This program will develop top notch researchers in the study of systemic autoimmune diseases who will lead basic and clinical research in academic centers to advance the care of people suffering from these diseases
Keywords: Autoimmune Status; Autoimmunity; Training Programs; self recognition (immune)
Project start date: 2008-08-01
Project end date: 2013-05-31
Budget start date: 1-JUN-2010
Budget end date: 31-MAY-2011
PFA/PA: PA-06-468
5T32AI074549-03 (2010): $123800
5T32AI074549-02 (2009): $124861
1T32AI074549-01A1 (2008): $61659
CIS School On Systemic Autoimmune Diseases
George C Tsokos, Professor Of Medicine
Clinical Immunology Society Suite 1100 Milwaukee, Wi 53202
Grant 1R13AR055043-01 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases IRG: AMS
Abstract: This grant requests support for travel and expenses of fellows to attend the 2007 Clinical Immunology Society (CIS) School on Systemic Autoimmune Diseases. This grant is requested to help defray travel costs for the U. S fellow travel and expenses to attend the School. Additional funding will be solicited from pharmaceutical and biotechnology companies. This School will be an intensive four-day residential course to be held in March 14-18, 2007 in Santa Fe, New Mexico. The program will be advertised through academic program directors and members of CIS, The American College of Rheumatology and American Association of Allergy and Applied Immunology as well as through other major societies with interests in clinical immunology. Advertising emphasis will be given to attract applicants representing minorities. The topic of the School will be Systemic Autoimmune Diseases, geared toward fellows in training holding either an MD an/or a PHD, within their last years of fellowship training. The primary goal of the school is education on the diagnosis, pathogenesis and treatment of systemic autoimmune diseases such as SLE, Scleroderma, Arthritis, and Vasculitis as well as gene therapy. These presentations will include topic overviews, state of the art establishes treatments, laboratory approaches, pathogenesis and future biologics. Emphasis will be given to new treatments that are under development and future possibilities. Sufficient time will be allocated for fellows to present either interesting cases or their work. By the end of the School, participants should be able to better diagnose and treat systemic autoimmune diseases, have an enhanced awareness of clinical immunology processes as they relate to systemic autoimmune diseases and its importance in scientific discoveries and clinical application; and become ambitious in pursuing academic careers to further our understanding, diagnosis and treatment of human systemic autoimmune diseases. Lastly, the School should stimulate future collaborations between young investigators in different medical centers and countries, and between young investigators and experienced physician/scientists in the field. The school will be residential in character and held in an informal setting and will attract applicants from the North America, Europe and Asia. Ten faculty and 25 fellows will be selected to attend.
Keywords: school, Asia, Europe, Mexico, advertising, art, arthritis, awareness, biotechnology, career, diagnosis, education, experience, gene therapy, health /scientific organization, human, hypersensitivity, immunology, motivation, physician, scleroderma, training, travel, university, vasculitis
Project start date: 2007-03-01
Project end date: 2007-08-31
1R13AR055043-01 (2007): $20000
2R13AR052254-02 (2006): $10000
1R13AR052254-01A1 (2005): $2000
REGULATION OF COMPLEMENT RECEPTOR GENE EXPRESSION
George C Tsokos, Professor Of Medicine
Henry M. Jackson Fdn For The Adv Mil/med
advancement Of Military Medicine, Inc.
rockville, Md 20852
Grant 5R01AI042782-03 from National Institute Of Allergy And Infectious Diseases IRG: GMA
Abstract: The proteins of the complement system participates in the regulation of various phases of the immune response. Complement activation products bind to immune complexes and form multivalent complement receptor 2 (CR2) ligands that regulate B cell proliferation and differentiation. The potential for using CR2 ligands to enhance antibody production urges the deciphering of the subcellular mechanisms that are involved in the regulation of transcription of the CR2 gene and the expression of its product on the surface membrane. We have identified a heterogeneous ribonucleoprotein (hnRNP) which binds to a previously unidentified DNA motif that is present in the promoter region of CR2 and is involved in the regulation of the transcription of the CR2 gene. On the basis of these findings we hypothesize that the hnRNP undergoes modification (phosphorylation) in response to external stimuli, binds to a DNA motif and acts in concert with other DNA binding proteins to control the transcription of the CR2 gene. The overall goal of this proposal is to complete the structural and functional characterization of the hnRNP protein and characterize the mechanisms that are involved in the regulation of the expression of complement CR2. Specifically we intend to structurally and functionally characterize the hnRNP that binds a CR2-promoter region-defined oligonucleotide and increases its transcriptional activity. This will be accomplished by determining the primary structure of hnRNP, by structurally and functionally analyzing the CR2-promoter region defined oligonucleotide binding and other functional domains of the hnRNP. We will characterize the mechanisms whereby external stimuli regulate the expression of CR2. This will be accomplished by investigating the effect of lymphokines on the transcription (CR2-promoter region defined oligonucleotide-driven reporter gene construct) and expression of the CR2 gene (mRNA and surface protein) and by characterizing the interactions of CR2 promoter region binding proteins. Since we have found that sera from patients with systemic autoimmune diseases have antibodies directed against this hnRNP, we propose to study their frequency and clinical significance
Keywords: complement pathway, complement receptor, gene expression, genetic transcription, heterogeneous nuclear ribonucleoprotein, receptor binding antinuclear autoantibody, autoimmune disorder, binding protein, binding site, genetic promoter element, lymphokine, protein binding, protein structure /function, transcription factor clinical research, epitope mapping, gel mobility shift assay, human subject
Project start date: 1998-12-01
Project end date: 2002-11-30
5R01AI042782-03 (2001): $161772
5R01AI042782-02 (2000): $157061
1R01AI042782-01A1 (1999): $152484
EXPANDED DOUBLE NEGATIVE T CELLS IN SLE
George C Tsokos, Professor Of Medicine
Beth Israel Deaconess Medical Center, Boston, Ma 02215
Grant 1R01AI085567-01 from National Institute Of Allergy And Infectious Diseases
Abstract: Systemic lupus erythematosus afflicts 1 to 2 million Americans and is associated with significant morbidity and mortality. While wise use of steroids and cytotoxics drugs along with effective use of antibiotics overall survival and quality of life have increased significantly over the last 30 years, we still lack specific treatment, and among others, the lack of full understanding of involved pathogenic mechanisms and of proper disease biomarkers are to be blamed for this. Immune cell and cytokine abnormalities in patients with systemic lupus erythematosus (SLE) are diverse and involve among others decreased cytotoxic responses, decreased activation induced cell death, decreased T regulatory function, increased dendritic cell function, increased IFNa, IL-6 and decreased IL-2, IFN? and TNFa production. Distinct molecular and biochemical abnormalities may account for several cell and cytokine abnormalities. Among peripheral T cells an expanded population of T cells missing CD4 and CD8 from the surface (double negative (DN) T cells exists which when placed in coculture with autologous B cells promoted the production of immunoglobulin and anti-dsDNA. We have recently found that this DN T cell population in SLE patients can be expanded in vitro and that it produces IL- 17 and more importantly, IL-17 producing DN T cells infiltrated kidney tissue of patients with lupus nephritis. We propose, accordingly, that an expanded DN T cell population in patients with SLE produces IL-17 and participates in target organ damage. We will test our thesis in experiments outlined in this application and grouped in four specific aims In the first aim we will establish the presence of CD4+ and expanded DN T cells producing IL-17 in patients with SLE-determine clinical correlations. In the second we will determine the origin of DN T cells is SLE patients, the requirements for expansion, their early and late signaling profile and their susceptibility to cellular control mechanisms. In the third we will determine the requirements for the generation of TH17 in SLE. In the last aim we study the nature of T cells that infiltrate target tissues in SLE and explore mechanisms which are responsible for the inappropriate homing. These studies intend to propose IL-17 as a novel treatment target in SLE patients and will generate information which will propose Th17 cells as a biomarker of disease activity. Systemic lupus erythematosus afflicts more than one million Americans most of whom are women in the child bearing age. Diagnosis is frequently delayed and the disease has significant morbidity and mortality. Current treatment is based primarily on indiscriminate immunosuppression. This proposal will identify IL-17 as a novel treatment target and will generate information to introduce Th17 cells as a biomarker of disease activity
Keywords: Accounting; Age; Alferon; American; Antibiotic Agents; Antibiotic Drugs; Antibiotics; Attention; Autologous; B blood cells; B cell differentiation factor; B cell stimulating factor 2; B-Cell Differentiation Factor-2; B-Cell Stimulatory Factor-2; B-Cells; B-Lymphocytes; BCDF; BSF-2; BSF2; BSF2 (B cell stimulating factor 2); Biochemical; Body Tissues; Bursa-Dependent Lymphocytes; Bursa-Equivalent Lymphocyte; CD8; CD8B; CD8B1; CD8B1 gene; CTLA-8; CTLA8; Cell Communication and Signaling; Cell Death; Cell Function; Cell Line; Cell Lines, Strains; Cell Process; Cell Signaling; Cell physiology; CellLine; Cells; Cellular Function; Cellular Physiology; Cellular Process; Clinical; Co-Stimulator; Co-culture; Cocultivation; Coculture; Coculture Techniques; Costimulator; Cytotoxic T-Lymphocyte-Associated Antigen 8; Cytotoxic T-Lymphocyte-Associated Serine Esterase 8; Cytotoxic agent; Cytotoxic drug; Data; Dendritic Cells; Diagnosis; Differentiation Factor, B-Cell; Disease; Disorder; Epidermal Thymocyte Activating Factor; G-interferon; Generations; Ginterferon; HPGF; Hepatocyte-Stimulating Factor; Homing; Hybridoma Growth Factor; IFN; IFN Alpha; IFN-beta 2; IFNB2; IFNa; IL-17; IL-17A; IL-2; IL-6; IL17; IL17 Protein; IL17A; IL2; IL2 Protein; IL6 Protein; Immune; Immune Globulins; Immunoglobulins; Immunoglobulins / Antibodies; Immunosuppression Effect; Immunosuppressions (Physiology); Immunosuppressive Effect; In Vitro; Interferon Alfa-n3; Interferon, Leukocyte; Interferon, Lymphoblast; Interferon, Lymphoblastoid; Interferon-alpha; Interferons; Interleukin 17 (Cytotoxic T-Lymphocyte-Associated Serine Esterase 8); Interleukin 17 Precursor; Interleukin 2; Interleukin 2 Precursor; Interleukin 6 (Interferon, Beta 2); Interleukin II; Interleukin-17; Interleukin-2; Interleukin-6; Interleukine 2; Interleukine 2 Precursor; Interleukine II; Intracellular Communication and Signaling; Kidney; LYT3; Lupus Erythematosus Disseminatus; Lupus Erythematosus, Systemic; Lupus Glomerulonephritis; Lupus Nephritis; Lymphocyte Mitogenic Factor; MGI-2; MHC Receptor; Major Histocompatibility Complex Receptor; Miscellaneous Antibiotic; Mitogenic Factor; Molecular; Morbidity; Morbidity - disease rate; Mortality; Mortality Vital Statistics; Myeloid Differentiation-Inducing Protein; Natural immunosuppression; Nature; Organ; Patients; Peripheral; Plasmacytoma Growth Factor; Population; Predisposition; Production; QOL; Quality of life; Receptors, Antigen, T-Cell; SLE; SLE - Lupus Erythematosus, Systemic; Signal Transduction; Signal Transduction Systems; Signaling; Steroid Compound; Steroids; Subcellular Process; Surface; Susceptibility; Systemic Lupus Erythematosus; Systemic Lupus Erythmatosus; T cell growth factor; T-Cell Growth Factor; T-Cell Receptor; T-Cell Stimulating Factor; T-Cells; T-Lymphocyte; Testing; Thymocyte Stimulating Factor; Thymus-Dependent Lymphocytes; Tissues; Urinary System, Kidney; Veiled Cells; Woman; base; bear children; bearing children; biological signal transduction; biomarker; child bearing; childbearing; cultured cell line; cytokine; cytotoxic; disease/disorder; disseminated lupus erythematosus; experiment; experimental research; experimental study; immunosuppression; interferon beta 2; necrocytosis; novel; peripheral blood; public health relevance; renal; research study; response; systemic lupus erythematosis; therapeutic target; thymus derived lymphocyte
Relevance: Systemic lupus erythematosus afflicts more than one million Americans most of whom are women in the child bearing age. Diagnosis is frequently delayed and the disease has significant morbidity and mortality. Current treatment is based primarily on indiscriminate immunosuppression. This proposal will identify IL-17 as a novel treatment target and will generate information to introduce Th17 cells as a biomarker of disease activity
Project start date: 2010-05-01
Project end date: 2015-04-30
Budget start date: 1-MAY-2010
Budget end date: 30-APR-2011
PFA/PA: PA-07-070
1R01AI085567-01 (2010): $434479
Sponsored Links Excellgen http://Excellgen.com
LYMPHOCYTE SIGNALING DEFECTS IN PATIENTS WITH LUPUS
George C Tsokos, Professor Of Medicine
Beth Israel Deaconess Medical Center, Boston, Ma 02215
Grant 5R01AI042269-14 from National Institute Of Allergy And Infectious Diseases
Abstract: Systemic lupus erythematosus (SLE) is characterized by diverse clinical and laboratory findings and multiple, frequently opposing cellular and cytokine aberrations. The T cell receptor (TCR) 6 chain is decreased in SLE T patients because of limited promoter activity, instability of an alternatively spliced TCR 6 mRNA missing crucial AU response elements and increased degradation by caspase 3. Its pace in the CD3 complex is taken by the FcR3 chain which signals through Syk. Lipid rafts form aggregates on the surface of T cells and together with the rewired TCR account for the aberrant signaling process in SLE T cells. In the lipid aggregates the adhesion molecule is present in increased amounts, which signal through ezrin. CD44 and phosphorylated ezrin positive cells are found in the kidneys of patients with lupus nephritis. It is hypothesized that altered cell surface membrane-mediated signaling in SLE T cells results in aberrant T cell function including inappropriate expression of adhesion molecules and homing to inflamed tissues. The objective of the proposed studies is to explore the role of TCR rewiring (FcR3 -> CD36) in effector SLE T cell function and establish the central role of Syk-mediated signaling in the expression of the SLE T cell phenotype and disease pathology. In addition, the contribution of cell membrane-mediated signaling in upregulating the expression of the adhesion molecule CD44 that results in inappropriate homing of T cells to the kidney will be studied. The proposed studies suggest the presence of molecular targets that can be modulated with gene transfer or drugs and have the potential to complement existing therapeutic modalities. In addition, they will identify several potential T cell based biomarkers. The long term goal of the ongoing and proposed research is the establishment of a "molecular signature" for each patient, or subgroups of SLE patients that will help the diagnosis and personalized treatment. Systemic lupus erythematosus afflicts more than one million Americans most of whom are women in the child bearing age. Current treatment is based primarily on indiscriminate immunosuppression. This application will explore the presence of molecular targets that can be modulated with gene transfer or drugs and have the potential to complement existing therapeutic modalities
Keywords: 3` Untranslated Regions; 3`UTR; Accounting; Adhesion Molecule; Age; American; Anti CD3; Anti-CD3 Antibody; Apopain; Behavior; Binding; Binding (Molecular Function); Biochemical; Body Tissues; CASP-3; CASP3; CD3; CD3 Antigens; CD3 Complex; CD3 molecule; CD36; CD36 gene; CD44; CD44 gene; CPP-32; CPP32; CPP32 protein; CPP32B; CPP32beta; CRE; CXCL12; CXCL12 gene; Caspase 3, Apoptosis-Related Cysteine Protease; Cell Adhesion Molecules; Cell Communication and Signaling; Cell Function; Cell Process; Cell Signaling; Cell membrane; Cell physiology; Cell surface; Cells; Cellular Function; Cellular Physiology; Cellular Process; Clinical; Complement; Complement Proteins; Complex; Cyclic AMP Response Element; Cysteine Protease CPP32; Cytokines, Chemotactic; Cytoplasmic Membrane; Defect; Diagnosis; Drugs; Enhancers; Expression Profiling; Expression Signature; GP3B; GP4; GPIV; Gene Transfer; Goals; HUM291 [Anti-CD3 Antibody]; Homing; Homologous Chemotactic Cytokines; HuM291; Immunosuppression Effect; Immunosuppressions (Physiology); Immunosuppressive Effect; Intercrines; Intracellular Communication and Signaling; Kidney; Lab Findings; Laboratory Finding; Lead; Lipid Rafts, Cell Membrane; Lipids; Lupus; Lupus Erythematosus Disseminatus; Lupus Erythematosus, Systemic; Lupus Glomerulonephritis; Lupus Nephritis; Lymphocyte; Lymphocytic; MDU3; MHC Receptor; Major Histocompatibility Complex Receptor; Mediating; Medication; Membrane; Membrane Microdomains; Messenger RNA; MoAb HuM291; Modality; Molecular; Molecular Fingerprinting; Molecular Interaction; Molecular Profiling; Molecular Target; Monoclonal Antibody HuM291; Natural immunosuppression; OKT3 antigen; PARP Cleavage Protease; PBSF; Pathology; Patients; Pb element; Peripheral; Pgp1; Pharmaceutic Preparations; Pharmaceutical Preparations; Phosphorylation; Plasma Membrane; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Protein Phosphorylation; RNA Splicing; RNA, Messenger; Receptors, Antigen, T-Cell; Relative; Relative (related person); Research; Response Elements; Rho-associated kinase; Rho-kinase; Role; SCA-1; SCARB3; SCYB12; SDF-1A; SDF-1B; SDF1; SDF1A; SDF1B; SIS cytokines; SLE; SLE - Lupus Erythematosus, Systemic; SREBP Cleavage Activity 1; SUBGP; Signal Transduction; Signal Transduction Systems; Signaling; Signaling Molecule; Sphingolipid Microdomains; Sphingolipid-Cholesterol Rafts; Splicing; Subcellular Process; Subgroup; Surface; Systemic Lupus Erythematosus; Systemic Lupus Erythmatosus; T-Cell Receptor; T-Cells; T-Lymphocyte; T3 Antigens; T3 Complex; T3 molecule; TLSF-A; TLSF-B; TPAR1; Therapeutic; Thymus-Dependent Lymphocytes; Tissues; Urinary System, Kidney; Woman; Yama; Yama protein; base; bear children; bearing children; biological signal transduction; biomarker; cAMP Response Element; caspase-3; cell adhesion protein; chemoattractant cytokine; chemokine; child bearing; childbearing; computerized data processing; cysteine protease P32; cytokine; data processing; disease phenotype; disseminated lupus erythematosus; drug/agent; experiment; experimental research; experimental study; ezrin; heavy metal Pb; heavy metal lead; immunosuppression; lipid raft; lymph cell; mRNA; membrane structure; membrane-organizing extension spike protein; moesin; molecuar profile; molecular signature; phosphoprotein p81; plasmalemma; public health relevance; radixin; radixin protein; receptor-mediated signaling; renal; research study; response; signal processing; social role; systemic lupus erythematosis; therapeutic target; thymus derived lymphocyte; transfer of a gene
Project start date: 1998-07-15
Project end date: 2013-05-31
Budget start date: 1-JUN-2010
Budget end date: 31-MAY-2011
PFA/PA: PA-07-070
5R01AI042269-14 (2010): $420750
5R01AI042269-13 (2009): $425000
2R01AI042269-12A1 (2008): $425000
2R01AI042269-06 (2003): $187125
5R01AI042269-04 (2001): $264902
5R01AI042269-03 (2000): $257185
5R01AI042269-02 (1999): $249689
1R01AI042269-01A1 (1998): $260265
George C Tsokos, Professor Of Medicine
Beth Israel Deaconess Medical Center, Boston, Ma 02215
Grant 5R01AI068787-04 from National Institute Of Allergy And Infectious Diseases
Abstract: Systemic lupus erythematosus (SLE) is an autoimmune disorder of indeterminate etiology characterized by profound T cell effector dysfunction. T cells from SLE patients produce decreased amounts of interleukin 2 (IL-2) in response to antigen stimulation. Sufficient amounts of IL-2 are needed to generate effector cytotoxic and T regulatory cells and for the proper elimination of autoreactive T cells through activation-induced cell death. Because the levels of IL-2 are determined at the IL-2 gene transcription level, we have designed studies to understand the involved mechanisms that lead to transcriptional repression of the IL-2 gene and devise approaches to correct it. We have observed that SLE T cells have increased protein and mRNA levels of the transcriptional repressor, cAMP responsive element (CRE) modulator (CREM), which binds to the -180 (-164/-189) site of the IL-2 promoter and suppress the IL-2 gene transcription. At the same site of the promoter binds the enhancer phosphorylated CRE binding protein (pCREB) and we hypothesize that the ratio of pCREB/CREM determines the transcriptional activity of the IL-2 promoter. We recently observed that a serine/threonine protein phosphatase (PP)2A is aberrantly expressed in SLE T cells and that it dephosphorylates pCREB and therefore, tilts the balance of pCREB/CREM bound at the -180 site of the IL-2 promoter towards CREM. The hypothesis will be tested in three sets of experiments. In the first, it will be established that PP2A is expressed at increased levels in patients with SLE, in the second, the mechanisms whereby increased PP2A activity causes decreased IL-2 production will be deciphered, whereas in the third, the mechanisms that are responsible for the increased expression of PP2Ac in SLE T cells will be explored. The generated data will shed light on to the molecular origin of decreased IL-2 production in SLE T cells that is claimed to be responsible for the decreased generation of cytotoxic T cells and increased rate of infections, the decreased numbers of T regulatory cells and the defective antigen activation-induced T cell death. In addition, the produced information will introduce new approaches to correct the abnormal expression of PP2Ac and reverse the decreased IL-2 production and defective T cell function
Keywords: AP-1; AP-1 Enhancer-Binding Protein; AP1; AP1 protein; ATGN; Activator Protein-1; Address; Aldesleukin Gene; Antigen Receptors; Antigens; Antimorphic mutation; Autoimmune Diseases; Binding; Binding (Molecular Function); Binding Proteins; Blood Serum; CRE Binding Protein; CREB; CREB Protein; CREM protein; CTL; Causality; Cell Communication Process; Cell Communication and Signaling; Cell Death; Cell Function; Cell Line; Cell Lines, Strains; Cell Process; Cell Signaling; Cell Signaling Process; Cell physiology; Cell-Mediated Lympholytic Cells; CellLine; Cells; Cellular Function; Cellular Oncogene c-FOS; Cellular Physiology; Cellular Process; Co-Stimulator; Costimulator; Cyclic AMP Response Element Modulator; Cyclic AMP Response Element-Binding Protein; Cyclic AMP Responsive Element Binding Protein; Cyclic AMP-Responsive DNA-Binding Protein; Cytolytic T-Cell; Cytotoxic T Cell; Cytotoxic T-Lymphocytes; Data; Defect; Disease; Disorder; Distal; Dominant Negative; Dominant-Negative Mutant; Dominant-Negative Mutation; Drugs; Dysfunction; Enhancer-Binding Protein AP1; Enhancers; Epidermal Thymocyte Activating Factor; Equilibrium; Etiology; Event; FBJ Murine Osteosarcoma Viral Oncogene Homolog; FOS Family Protein; FOS Oncoprotein; FOS Protein; FOS gene; Functional disorder; G0S7; G0S7 Protein; Gene Down-Regulation; Gene Transcription; Generations; Genes; Genetic Transcription; Hour; Human; Human, General; ICER protein; ICERI protein; IL-2; IL-2 Gene; IL2; IL2 Protein; IL2 gene; Immunosuppressants; Immunosuppressive Agents; Infection; Interleukin 2; Interleukin 2 Precursor; Interleukin 2 Precursor Gene; Interleukin II; Interleukin-2; Interleukin-2 Gene; Interleukine 2; Interleukine 2 Precursor; Interleukine II; Intracellular Communication and Signaling; L-Serine; L-Threonine; Lead; Ligand Binding Protein; Light; Lupus; Lupus Erythematosus Disseminatus; Lupus Erythematosus, Systemic; Lymphocyte Mediators; Lymphocyte Mitogenic Factor; Lymphokines; Man (Taxonomy); Man, Modern; Mediating; Medication; Membrane; Messenger RNA; Mitogenic Factor; Molecular; Molecular Interaction; Morbidity; Morbidity - disease rate; Mortality; Mortality Vital Statistics; Outcome; PP2A; PP2A Subunit B Prime; PPP2R4; PR53; PTPA; Pathogenesis; Patients; Pb element; Pharmaceutic Preparations; Pharmaceutical Preparations; Phosphatases; Phosphohydrolases; Phosphomonoesterases; Phosphoprotein Phosphatase; Phosphoprotein Phosphatase-2C; Phosphoprotein Phosphohydrolase; Phosphoric Monoester Hydrolases; Phosphotyrosyl Phosphatase Activator; Photoradiation; Physiopathology; Post-Transcriptional Gene Silencing; Post-Transcriptional Gene Silencings; Posttranscriptional Gene Silencing; Posttranscriptional Gene Silencings; Production; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Protein Binding; Protein Phosphatase 2A Regulatory Subunit B Prime; Protein Phosphatase 2A Regulatory Subunit PR53; Protein Phosphatase C; Protein Phosphatase-1; Protein Phosphatase-2A; Protein Serine/Threonine Phosphatase; Protein phosphatase; Proteins; Proto-Oncogene Products c-fos; Proto-Oncogene Proteins c-fos; Protooncogene FOS; Quelling; RNA Expression; RNA Interference; RNA Silencing; RNA Silencings; RNA, Messenger; RNA, Small Interfering; RNAi; Receptor Protein; Receptors, Antigen; Regulatory T-Lymphocyte; Research Design; SLE; SLE - Lupus Erythematosus, Systemic; Sequence-Specific Posttranscriptional Gene Silencing; Serine; Serum; Signal Transduction; Signal Transduction Systems; Signaling; Site; Small Interfering RNA; Study Type; Subcellular Process; Subcellular Signaling Process; Systemic Lupus Erythematosus; Systemic Lupus Erythmatosus; T cell growth factor; T-Cell Growth Factor; T-Cell Growth Factor Gene; T-Cell Stimulating Factor; T-Cells; T-Lymphocyte; T-Lymphocyte, Regulatory; T-Lymphocytes, Cytotoxic; TCGF Gene; Testing; Threonine; Thymocyte Stimulating Factor; Thymus-Dependent Lymphocytes; Transcription; Transcription Corepressor; Transcription Factor AP-1; Transcription Regulation; Transcription Repression; Transcription Repressor; Transcription Repressor/Corepressor; Transcription, Genetic; Transcriptional Control; Transcriptional Corepressor; Transcriptional Regulation; Transcriptional Repression; Transcriptional Repressor; Transcriptional Repressor/Corepressor; autoimmune disorder; autoreactive T cell; balance; balance function; biological signal transduction; c fos; c-fos Gene; c-fos Protein; c-fos Proteins; c-fos Proto-Oncogenes; cAMP Response Element-Binding Protein; cAMP Responsive Element Binding Protein; cAMP-responsive element modulator; cultured cell line; cytotoxic; disease causation; disease etiology; disease/disorder; disease/disorder etiology; disorder etiology; disseminated lupus erythematosus; drug/agent; experiment; experimental research; experimental study; fos Proto-Oncogene Proteins; gene product; gene repression; heavy metal Pb; heavy metal lead; immunogen; immunosuppressive; mRNA; membrane structure; necrocytosis; new approaches; new diagnostics; new therapeutics; next generation diagnostics; next generation therapeutics; novel approaches; novel diagnostics; novel strategies; novel strategy; novel therapeutics; p55(c-fos); pathophysiology; receptor; research study; response; serine-threonine phosphatase; siRNA; study design; systemic lupus erythematosis; thymus derived lymphocyte; tool; transcription factor; v-FOS FBJ Murine Osteosarcoma Viral Oncogene Homolog
Project start date: 2007-04-01
Project end date: 2012-03-31
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
5R01AI068787-04 (2010): $405726
5R01AI068787-03 (2009): $409824
Sponsored Links Excellgen http://Excellgen.com
George C Tsokos, Professor Of Medicine
Beth Israel Deaconess Medical Center 330 Brookline Avenue, Br 264 Boston, Ma 02215
Grant 7R01AI049954-06 from National Institute Of Allergy And Infectious Diseases IRG: GMA
Abstract: Systemic lupus erythematosus (SLE) is an autoimmune disorder of indeterminate etiology characterized by profound T cell effector dysfunctions including defective interleukin-2 (IL-2) production. The fundamental underlying molecular mechanisms remain largely unknown. This laboratory has considered that deficient production of IL-2 represents decreased transcription of the IL-2 gene and has undertaken a systematic analysis to investigate aberrations of transcription factors that bind to the IL-2 promoter and cause defective IL-2 gene transcription. The presented preliminary studies show that IL-2 production by SLE T cells is decreased and the activity of two activators, nuclear factor (NF)-kappaB and activating protein (AP)-1, is decreased, whereas the activity of the repressor cAMP responsive element modulator (CREM) is increased. The latter binds to the -180 (-164/-189) site of the IL-2 promoter and suppresses gene transcription. This information suggests that the decreased IL-2 production in human SLE is the result of transcriptional repression mediated by the increased expression of CREM and/or the decreased activity of activators. This hypothesis will be addressed logically in experiments planned in 3 specific aims, which will 1) Establish in cross-sectional and longitudinal studies that the increased expression of CREM in human SLE subjects is disease-specific and disease activity-independent and that it relates directly with the decreased production of IL-2. 2) Determine how CREM suppresses the production of IL-2 in human SLE T cells by studying its ability to interact with other transcription factors on the IL-2 promoter. 3) Investigate the mechanisms that are involved in the increased expression of CREM in SLE T cells. Defective IL-2 production contributes to increased infection-related morbidity and mortality rates in patients with SLE. The proposed studies will contribute to our understanding of the origin of a major clinical problem.
Keywords: T lymphocyte, genetic transcription, immunogenetics, interleukin 2, molecular pathology, systemic lupus erythematosus, anergy, messenger RNA, transcription factor, human subject, patient oriented research, tissue /cell culture, transfection
Project start date: 2002-04-15
Project end date: 2007-12-31
1R01AI049954-01A1 (2002): $259350
CIS School In Systemic Autoimmune Diseases
George C Tsokos, Professor Of Medicine
Clinical Immunology Society
Grant 1R13AR057218-01 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases IRG: AMS
Abstract: SUMMARY This grant requests support for travel and expenses of fellows to attend the 2009, 2010, and 2011 Clinical Immunology Society (CIS) School in Systemic Autoimmune Diseases. This grant is requested to help defray costs for U.S. fellow travel and expenses related to attending the School. Additional funding will be solicited from pharmaceutical and biotechnology companies. The next three programs are scheduled to take place during the spring in Boston, Massachusetts. This activity is intended to reach an international target audience consisting of fellows-in-training from America, Europe, and Asia holding either an MD and/or a PhD and within their last years of fellowship training. Ten faculty and twenty-five fellows will be selected to attend each School. The program will be advertised through academic program directors and members of the CIS and the American College of Rheumatology as well as through other major societies with interests in clinical immunology. Advertising emphasis will be given to attract applicants representing minorities. The objective of the School is to enhance the ability of the attendants to diagnose and treat systemic autoimmune diseases; to improve their understanding of immunopathogenic processes as they relate to systemic autoimmune diseases; to familiarize with the importance of scientific discoveries as they translate into clinical practice; to promote ambition among young fellows to pursue academic careers in the study of systemic autoimmune diseases. Lastly, the School is designed to stimulate future collaborations between young investigators in different medical centers and countries, and between young investigators and experienced physician/scientists in the field
Project start date: 2009-04-01
Project end date: 2012-03-31
T Cell Help In The B Cell Response To Malaria CS Protein
George C Tsokos, Professor Of Medicine
Henry M. Jackson Fdn For The Adv Mil/med Advancement Of Military Medicine, Inc. Rockville, Md 20852
Grant 5R03AI053463-02 from National Institute Of Allergy And Infectious Diseases IRG: TMP
Abstract: Although malaria has been eradicated in most of the western world, it has become one of the most important emerging diseases today due to the increased resistance both of the parasite to drugs and the mosquito to insecticides. There has been tremendous effort to develop a vaccine to prevent the disease, but progress has been severely hampered by the lack of knowledge of how protective immunity is elicited against the parasite. Recent work suggests that the humoral immune response to malaria sporozoite antigens is driven along both T-dependent and T-independent pathways. However, the role of T cell help in stimulating protective immunity against sporozoites is controversial. The goal of this proposal is to produce the first comprehensive analysis of the humoral immune response to malaria sporozoite infection, and to determine the role of T-dependent and T-independent activation pathways in shaping the repertoire of antibodies utilized in the response. These goals will be addressed by examining the primary immune response to malaria sporozoite infection in normal and T cell-deficient mice. The response will be measured by first constructing a series of hybridoma libraries specific for the sporozoite s immunodominant antigen from infected normal and nude mice. Each of the libraries will be analyzed for the antibody repertoire utilized against the antigen in regards to recurrent clonotypes, isotype usage, fine specificity, and affinity maturation. Finally, the immune responses between libraries derived under T-dependent and T-independent conditions will be compared to determine what qualitative differences lay between them. This project will act as a pilot study for a later, more comprehensive study to delineate the role of T-dependent and T-independent activation pathways in provoking long-lived, protective immunity against malaria infection.
Keywords: B lymphocyte, T lymphocyte, antigen antibody reaction, cell cell interaction, circumsporozoite protein, humoral immunity, leukocyte activation /transformation, malaria vaccine, antigen presentation, cellular immunity, immunologic memory, malaria, vaccine development, athymic mouse, genetically modified animal, hybridoma, laboratory mouse, polymerase chain reaction, protein sequence
Project start date: 2003-07-01
Project end date: 2005-06-30
5R03AI053463-02 (2004): $74350
1R03AI053463-01A1 (2003): $74350
George C Tsokos
Beth Israel Deaconess Medical Center
Project start date: 2006-03-01
Project end date: 2017-03-31
George C Tsokos, Professor Of Medicine
Beth Israel Deaconess Medical Center 330 Brookline Avenue, Br 264 Boston, Ma 02215
Grant 1R01AI068787-01A1 from National Institute Of Allergy And Infectious Diseases IRG: ZRG1
Keywords: cell, human, RNA, antigen, antigen receptor, autoimmune disorder, balance, binding protein, cell death, cell line, disease /disorder etiology, element, experimental design, fos protein, gene, human subject, immunosuppressive, infection, interleukin 2, lead, lighting, lymphokine, membrane, phosphoprotein phosphatase, protein, protein binding, receptor, repression, sectioning, serine, serum, suppression, systemic lupus erythematosus, threonine, transcription factor, clinical research
Project start date: 2007-04-01
Project end date: 2012-03-31
1R01AI068787-01A1 (2007): $350261
Lymphocyte Signaling Defects In Patients With Lupus
George C Tsokos, Professor Of Medicine
Beth Israel Deaconess Medical Center 330 Brookline Avenue, Br 264 Boston, Ma 02215
Grant 5R01AI042269-11 from National Institute Of Allergy And Infectious Diseases IRG: ZRG1
Project start date: 1998-07-15
Project end date: 2008-12-31
5R01AI042269-11 (2007): $402977
Sponsored Links Excellgen http://Excellgen.com
5R01AI042269-09 (2006): $365455
5R01AI042269-08 (2005): $374250
5R01AI042269-07 (2004): $374250